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Green synthesis of nanoparticles by using plants is an emerging class of nanobiotechnology. It revolutionizes all the fields of nanobiotechnology by synthesizing chemical‐free nanoparticles for various purposes. In the present study, zinc and copper nanoparticles were synthesized by using the white leaves of Allium cepa and further characterized through Zeta analyzer and Scanning electron microscopy. Zeta analyzer elucidated that zinc nanoparticles ranged from 8‐32 nm while copper nanoparticles ranged from 15‐30 nm. Scanning electron microscopy clarified that zinc nanoparticles were irregular while copper nanoparticles were spherical in shape. The effects of green synthesized nanoparticles were evaluated on the germination frequency and biochemical parameters of plant tissues. The nucellus tissues were inoculated on Murashige and Skoog (MS) medium augmented with 30 µg/ml suspension of zinc and copper nanoparticles. Green synthesized nanoparticles enhanced the in vitro germination parameters because of their low toxicity and high efficacy. Significant results were obtained for germination parameters in response to the applications of zinc nanoparticles as compared to copper nanoparticles. These nanoparticles could also induce stress in plantlets by manipulating the endogenous mechanism as a result various defence compounds are produced which have potential in treating various human ailments. Copper nanoparticles showed higher toxicity as compared to zinc nanoparticles and triggered the production of antioxidative enzymes and non‐ enzymatic compounds. [ABSTRACT FROM AUTHOR]

The globalization process during last two hundred years has influenced the world's social, cultural and economic positions. Consequently, world's output, trade and population have been increased significantly. As a result, wellbeing of nations has been improved. However, at the same time in equality among the rich and poor nations of the world has also increased which is evident from the "Gini coefficient" reported by the World Trade Organization (WTO), 2013. Therefore this study aims to investigate export determining factors for four neighboring countries namely China, India, Pakistan and Bangladesh. It is an empirical analysis based on the structural gravity model. The study employees a relatively new technique of panel data (PPML-Estimator) for the comparison between countries. The findings reveal that factors such as income, expenditure and geographical distance in all four countries are in line with the theoretical literature as well as consistent with gravity theory. However, the dummy variables for language, contiguity and the Regional Trade Agreement (RTAs) have mixed results. Notably, the RTAs in case of China, India and Pakistan have negative and significant impact. In another model, in order to capture the impact of globalization, this study used overall globalization index (GI) and its proxies like Political Globalization Index (PGI), Social Globalization Index (SGI), Economic Globalization Index (EGI) and Information Flow Index (IFI). The results suggest that the proxies have mixed results whereas; the overall globalization has positive and significant effect on the exports. On the basis of these finding, it may be concluded that political conflicts among these neighboring nations are the main hurdle for their mutual beneficial trade. Although, Pakistan's socio-economic set up is responsive towards globalization wave however, factors like poor management, energy crises, corruption and terrorism have hampered its trade performance. [ABSTRACT FROM AUTHOR]

Background: The spread of artemisinin resistance in the Greater Mekong Subregion of Southeast Asia poses a significant threat for current anti-malarial treatment guidelines globally. The aim of this study was to assess the current prevalence of molecular markers of drug resistance in Plasmodium falciparum in the four provinces with the highest malaria burden in Pakistan, after introducing artemether–lumefantrine as first-line treatment in 2017. Methods: Samples were collected during routine malaria surveillance in Punjab, Sindh, Baluchistan, and Khyber Pakhtunkhwa provinces of Pakistan between January 2018 and February 2019. Plasmodium falciparum infections were confirmed by rapid diagnostic test or microscopy. Plasmodium falciparum positive isolates (n = 179) were screened by Sanger sequencing for single nucleotide polymorphisms (SNPs) in the P. falciparum kelch 13 (pfk13) propeller domain and in P. falciparum coronin (pfcoronin). SNPs in P. falciparum multidrug resistance 1 (pfmdr1) N86Y, Y184F, D1246Y and P. falciparum chloroquine resistance transporter (pfcrt) K76T were genotyped by PCR-restriction fragment length polymorphism. Results: No artemisinin resistance associated SNPs were identified in the pfk13 propeller domain or in pfcoronin. The pfmdr1 N86, 184F, D1246 and pfcrt K76 alleles associated with reduced lumefantrine sensitivity were present in 83.8% (150/179), 16.9% (29/172), 100.0% (173/173), and 8.4% (15/179) of all infections, respectively. The chloroquine resistance associated pfcrt 76T allele was present in 98.3% (176/179) of infections. Conclusion: This study provides an update on the current prevalence of molecular markers associated with reduced P. falciparum sensitivity to artemether and/or lumefantrine in Pakistan, including a first baseline assessment of polymorphisms in pfcoronin. No mutations associated with artemisinin resistance were observed in pfk13 or pfcoronin. However, the prevalence of the pfmdr1 N86 and D1246 alleles, that have been associated with decreased susceptibility to lumefantrine, remain high. Although clinical and molecular data suggest that the current malaria treatment guidelines for P. falciparum are presently effective in Pakistan, close monitoring for artemisinin and lumefantrine resistance will be critical to ensure early detection and enhanced containment of emerging ACT resistance spreading across from Southeast Asia. [ABSTRACT FROM AUTHOR]

For nanovaccine-based cancer immunotherapy, dendritic cells (DCs) are one of the most powerful antigen presenting cells (APCs) that initiate and promote the maturation of antigen-specific cytotoxic T lymphocytes ( e.g. , CD8 + T cells) to induce the local and systemic antitumor immunity and further suppress the tumor metastasis and produce long-term protection against tumor. Thus, the activation and maturation of DCs is the prerequisite for efficient CD8 + T cell-based antitumor immune responses, which is considered as a primary and promising task for nanovaccine engineering. Herein, we introduce a versatile nanovaccine of liposomes-coated gold nanocages (Lipos-AuNCs) modified with DCs specific antibody aCD11c for targeted delivery of adjuvant MPLA and melanoma antigen peptide TRP2 to promote the activation and maturation of DCs, and enhance tumor specific T lymphocytes responses. Moreover, AuNCs accumulation and AuNCs-engulfed DCs migration to regional lymph nodes (RLNs) became real-time visualization through in vivo fluorescence and photoacoustic (PA) imaging to monitor the immunity process. In vivo experimental results demonstrated that the targeted antigen/adjuvants-loaded AuNCs exhibited enhanced antitumor immune response to inhibit tumor growth and metastasis in both B16-F10 prophylactic and lung metastasis models, which may act as a promising nanoplatform for antitumor immunotherapy and in vivo tracking. [ABSTRACT FROM AUTHOR]

Glucose-6-phosphate dehydrogenase (G6PD) is a key enzyme in the pentose phosphate pathway that ensures sufficient production of coenzyme nicotinamide adenine dinucleotide phosphate (NADPH) by catalyzing the reduction of NADP+ to NADPH. Noteworthy, the latter mediates the production of reactive oxygen species (ROS) by phagocytic cells such as neutrophils and monocytes. Therefore, patients with severe forms of G6PD deficiency may present impaired NADPH oxidase activity and become susceptible to recurrent infections. This fact, highlights the importance to characterize the immunopathologic mechanisms underlying the susceptibility to infections in patients with G6PD deficiency. Here we report the first two cases of G6PD deficiency with Bacille Calmette-Guérin (BCG) adverse effect, besides jaundice, hemolytic anemia and recurrent infections caused by Staphylococcus aureus . The qualitative G6PD screening was performed and followed by oxidative burst analysis using flow cytometry. Genetic and in silico analyses were carried out by Sanger sequencing and mutation pathogenicity predicted using bioinformatics tools, respectively. Activated neutrophils and monocytes from patients displayed impaired oxidative burst. The genetic analysis revealed the novel missense mutation c.1157T>A/p.L386Q in G6PD. In addition, in silico analysis indicated that this mutation is pathogenic, thereby hampering the oxidative burst of neutrophils and monocytes from patients. Our data expand the clinical and genetic spectrum of G6PD deficiency, and suggest that impaired oxidative burst in this severe primary immune deficiency is an underlying immunopathologic mechanism that predisposes to mycobacterial infections. [ABSTRACT FROM AUTHOR]

Chronic granulomatous disease (CGD) is a primary immunodeficiency caused by mutations in the five structural genes (CYBB , CYBA, NCF1, NCF2 , and NCF4) that typically results in a decrease in function or inability to generate a respiratory burst, leading to defective killing of pathogens, including fungi and intracellular bacteria. Mutations in CYBB , encoding the gp91phox (also known as NOX2) result in X-linked CGD account for approximately 65% of CGD cases. Here, we aimed the characterization of a novel missense mutation c.1226C > A/p.A409E in the CYBB gene in a patient with X-linked CGD. Relevant clinical data of a male patient whose family was positive for XCGD was reviewed. Oxidative burst and NADPH protein expression was evaluated by flow cytometry, while Genetic analysis was performed by Sanger sequencing. Monocyte-derived macrophages (MDMs) were evaluated for their capacity for phagocytosis and growth suppression of the intracellular Mycobacterium tuberculosis ( M. tuberculosis ). We thus report the absence of an oxidative burst in the phagocytes of the patient. Flow cytometry evaluation revealed a normal expression of NADPH oxidase components in neutrophils and genetic analysis proved the existence of a novel missense c.1226C > A mutation in the CYBB gene resulting in p.A409E. Further, we have showed that the patient's MDMs were unhindered in their ability to take up mycobacteria normally. Instead, the MDMs failed to control the intracellular proliferation of M. tuberculosis , a phenotype that improved in the presence of recombinant human interferon-gamma (rhIFN-γ). This work expands the genetic spectrum of X-linked CGD and demonstrates improvement in macrophage function in X91 + CGD patient by rhIFN-γ. [ABSTRACT FROM AUTHOR]

Tuberculosis is a global health problem, and its early diagnosis is the ultimate strategy for prevention and control. The current study was undertaken to evaluate conventional and molecular diagnostic assays for the detection of mycobacteria in pulmonary tuberculosis (TB) patients from Khyber Pakhtunkhwa region of Pakistan. A total of 259 clinically suspected patients of TB were processed for Zeihl Neelsen (ZN) microscopy, BACTEC MGIT liquid culture and GeneXpert assay. Among 259 samples, 28 (10.81%) were positive for acid fast bacilli (AFB) on ZN microscopy. In liquid culture, the growth of mycobacterium species was obtained in 36 (13.89%) samples while the GeneXpert assay detected Mycobacterium tuberculosis (MTB) in 49 (18.91%) samples. Detection rate of MTB was significantly high (n = 49, p < 0.0095) on GeneXpert as compared to microscopy (n = 28); however no significant difference (p = 0.1230) was observed on GeneXpert (n = 49) and culture (n = 36) based detection of MTB. The strength of agreement between GeneXpert and microscopy was also poor (Kappa value < 0.114, 95% CI: −0.72 – 0.301) which support our results. MTB detection rate among female was high as compared to male TB patients while in age wise, the age group 55–64 years has almost high detection rate on microscopy, culture and GeneXpert assay. Findings of the present study highlighted that GeneXpert is more efficient tool for timely diagnosis and proper TB control in high TB endemic area. [ABSTRACT FROM AUTHOR]

Rapid identification of pathogens with higher sensitivity and specificity plays a significant role in maintaining public health, environmental monitoring, controlling food quality, and clinical diagnostics. Different methods have been widely used in food testing laboratories, quality control departments in food companies, hospitals, and clinical settings to identify pathogens. Some limitations in current pathogens detection methods are time-consuming, expensive, and laborious sample preparation, making it unsuitable for rapid detection. Microfluidics has emerged as a promising technology for biosensing applications due to its ability to precisely manipulate small volumes of fluids. Microfluidics platforms combined with spectroscopic techniques are capable of developing miniaturized devices that can detect and quantify pathogenic samples. The review focuses on the advancements in microfluidic devices integrated with spectroscopic methods for detecting bacterial microbes over the past five years. The review is based on several spectroscopic techniques, including fluorescence detection, surface-enhanced Raman scattering, and dynamic light scattering methods coupled with microfluidic platforms. The key detection principles of different approaches were discussed and summarized. Finally, the future possible directions and challenges in microfluidic-based spectroscopy for isolating and detecting pathogens using the latest innovations were also discussed., Competing Interests: The authors declare no conflict of interest., (© 2024 Author(s).)

Pancreatic ductal adenocarcinoma (PDAC) is a highly lethal cancer often diagnosed at advanced stages, highlighting the urgent need for early detection strategies. This systematic review explores the potential of fecal and urinary biomarkers for early PDAC detection. A comprehensive search identified eight relevant studies investigating various biomarkers, including proteins, metabolites, microbial profiles, DNA mutations, and non-coding RNAs. Promising findings suggest that urinary biomarkers related to metabolic alterations, inflammatory processes, fecal microbiome profiles, and fecal miRNAs hold diagnostic potential even at early stages of PDAC. Combining biomarkers into panels may enhance diagnostic accuracy. Challenges such as validation in larger cohorts, standardization of protocols, and regulatory approval must be addressed for clinical translation. Despite these hurdles, non-invasive urinary and fecal biomarkers represent a promising avenue for improving PDAC outcomes through early detection., Competing Interests: The authors have declared that no competing interests exist., (Copyright © 2024, Arif et al.)

In the present investigation, the role of GS-AgNPs treatment in wheat plants was carried out in reducing heat stress with the aim of facilitating scientists on this topic. The effect of GS-AgNPs against heat stress has rarely been deliberated in wheat plants, and only a few studies have been established earlier in this scenario. This work illustrated the effect of GS-AgNPs on the regulation of carbohydrates metabolism, SOD, proteins, crude fibers, and minerals changes in wheat plants. Data were analysed using PCA analysis, correlation parameters, and normal probability distribution in PAST 3 software. The results indicated that heat stress alone caused severe changes in carbohydrates metabolism, SOD, proteins, crude fibers, and minerals immediately so that plants could not recover without foreign stabilizers such as GS-AgNPs. The application of GS-AgNPs increases the flux of carbohydrates metabolism, SOD, and proteins, including HSPs, crude fibers, and minerals, in wheat plants to reduce the effect of heat stress. The 50 mg/l concentration of GS-AgNPs has shown an increase in carbohydrates metabolism and SOD activity, while crude fibres have shown a significant enhancement at 100 mg/l of GS-AgNPs. The crude and true proteins were also shown pronounced increase in treatment to a concentration of 50 mg/l of GS-AgNPs. GS-AgNPs stimulated HSP production; most importantly, smHSP production was observed in the present results with other HSPs in wheat plants treated with a 50 mg/l concentration of GS-AgNPs. The mineral distribution was also regulated by the respective treatment of GS-AgNPs, and the highest amounts of Ca, P and Fe were found to be highest in wheat under heat stress. In general, we computed the expected model based on GS-AgNPs on the genes/factors that respond to heat stress and their potential role in mitigating heat stress in wheat. In addition, we discussed the prospective signalling pathway triggered by GS-AgNPs in wheat against heat stress. In the future, this work might be helpful in distinguishing the genetic variation due to GS-AgNPs in promoting tolerance in wheat against heat stress.Communicated by Ramaswamy H. Sarma.

Aim: The current research aims to design effective strategies to enhance the body's immune system against pathogenic bacteria. Methods: Skin commensals were isolated, identified and cultured in fish collagen peptides (FCPs). Results: After culturing in FCP, the skin commensals were used in a dose-dependent manner for Staphylococcus aureus in a dual-culture test, which showed significant growth inhibition of the pathogenic bacteria, which concluded that FCP induced the immune defense system of skin microbiota against pathogenic strains. Conclusion: Results have validated that fish collagen peptide plays a vital role in the growth of selected human skin flora and induces more defensive immunity against pathogenic S. aureus bacteria in dual-culture experimentation.

Pseudomonas aeruginosa is an opportunist pathogen responsible for causing several infections in the human body, especially in patients with weak immune systems. The proposed approach reports a novel pathogens detection system based on cultivating microdroplets and acquiring the scattered light signals from the incubated droplets using a microfluidic device. Initially, the microdroplets were generated and incubated to cultivate bacteria inside the microdroplets. The second part of the microfluidic chip is the detection module, embedded with three optical fibers to connect laser light and photosensors. The incubated droplets were reinjected in the detection module and passed through the laser light. The surrounding photosensors were arranged symmetrically at 45° to the flowing channel for acquiring the scattered light signal. The noise was removed from the acquired data, and time-domain waveform features were evaluated. The acquired features were trained using machine learning classifiers to classify P. aeruginosa. The k-nearest neighbors (KNN) showed superior classification performance with 95.6 % accuracy among other classifiers, including logistic regression (LR), support vector machines (SVM), and naïve Bayes (NB). The proposed research was performed to validate the method for pathogens detection with a concentration of 10 5 CFU/mL. The total duration of 6 h is required to test the sample, including five hours for droplets incubation and one hour for sample preparation and detection using light scattering module. The results indicate that acquiring the light scattering patterns from incubated droplets can detect P. aeruginosa using machine learning classification. The proposed system is anticipated to be helpful as a rapid device for diagnosing pathogenic infections., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

The rapid classification of micro-particles has a vast range of applications in biomedical sciences and technology. In the given study, a prototype has been developed for the rapid detection of particle size using multi-angle dynamic light scattering and a machine learning approach by applying a support vector machine. The device consisted of three major parts: a laser light, an assembly of twelve sensors, and a data acquisition system. The laser light with a wavelength of 660 nm was directed towards the prepared sample. The twelve different photosensors were arranged symmetrically surrounding the testing sample to acquire the scattered light. The position of the photosensor was based on the Mie scattering theory to detect the maximum light scattering. In this study, three different spherical microparticles with sizes of 1, 2, and 4 μm were analyzed for the classification. The real-time light scattering signals were collected from each sample for 30 min. The power spectrum feature was evaluated from the acquired waveforms, and then recursive feature elimination was utilized to filter the features with the highest correlation. The machine learning classifiers were trained using the features with optimum conditions and the classification accuracies were evaluated. The results showed higher classification accuracies of 94.41%, 94.20%, and 96.12% for the particle sizes of 1, 2, and 4 μm, respectively. The given method depicted an overall classification accuracy of 95.38%. The acquired results showed that the developed system can detect microparticles within the range of 1-4 μm, with detection limit of 0.025 mg/ml. Therefore, the current study validated the performance of the device, and the given technique can be further applied in clinical applications for the detection of microbial particles., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 He, Wang, Wang, Yu, Zhao, Li, Hussain and Liu.)

Background: Leishmaniasis is the second and fourth highest cause of mortality and morbidity respectively among all tropical diseases. Recurrence in the onset of leishmaniasis is a major problem that needs to be addressed to reduce the case fatality rate and ensure timely clinical intervention. Here we are investigating the association of risk factors with recurrent cutaneous leishmaniasis to address this issue., Material and Methods: Patients received by Nasser Ullah Khan Babar Hospital in Peshawar, Pakistan from March 2019 to July 2020 were enrolled in this study. Those patients who developed symptoms after completion of treatment were included in Group-A while those who had atypical scars like leishmaniasis but were negative for cutaneous leishmaniasis were included in the comparison group tagged as Group B. All those individuals who had completed six weeks of treatment for CL but had normal complete blood counts (CBC) were included to avoid other underlying immunological pathologies, while we excluded those participants who had co-morbidities like diabetes, liver disease, cardiac disease, and pregnant and lactating women through their history Association was tested between Group-A and Group-B with other explanatory variables through chi-square test. The regression model was proposed to determine the predictors., Result: A total of 48 participants of both sexes were included in the study with a mean age of 32.2 ± 15.10. The data suggest that females are overrepresented among the patients with recurrent leishmaniasis [21(53.8 %,); p = 0.07]. Compared to patients; healthy participants had a higher proportion of adults (19-59 years) versus adolescents (13-18 years) [26(66.7 %) vs 07(17.9), p = 0.004]. Multivariate logistic regression analysis shows that females are 2.1 times more prone to infections among cases as compared to healthy individuals [unadjusted OR 2.20, 95 % confidence interval (CI) 1.5-10.6, p = 0.02; adjusted OR 2.1, 95 % CI 1.50-10.69, p = 0.02]. We propose that patients receiving intradermal were less likely to be infected as compared to those receiving intralesional injections [unadjusted OR 0.07.0, 95 % confidence interval (CI) 1.18-3.37, p = 0.03; adjusted OR 0.06, 95 % CI 1.18-3.38, p = 0.03]., Conclusion: Old age (adults) and sex (females) were the strongest predictors to be associated with recurrent leishmaniasis. Similarly, the choice of intradermal as compared to intralesional injection and the prolonged treatment duration were strongly associated with greater chances of recurrence., Competing Interests: Conflict of interest None declared., (Copyright © 2022 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Detection of foodborne pathogens at an early stage is very important to control food quality and improve medical response. Rapid detection of foodborne pathogens with high sensitivity and specificity is becoming an urgent requirement in health safety, medical diagnostics, environmental safety, and controlling food quality. Despite the existing bacterial detection methods being reliable and widely used, these methods are time-consuming, expensive, and cumbersome. Therefore, researchers are trying to find new methods by integrating spectroscopy techniques with artificial intelligence and advanced materials. Within this progress report, advances in the detection of foodborne pathogens using spectroscopy techniques are discussed. This paper presents an overview of the progress and application of spectroscopy techniques for the detection of foodborne pathogens, particularly new trends in the past few years, including surface-enhanced Raman spectroscopy, surface plasmon resonance, fluorescence spectroscopy, multiangle laser light scattering, and imaging analysis. In addition, the applications of artificial intelligence, microfluidics, smartphone-based techniques, and advanced materials related to spectroscopy for the detection of bacterial pathogens are discussed. Finally, we conclude and discuss possible research prospects in aspects of spectroscopy techniques for the identification and classification of pathogens.

Tick infestation is a major public and animal health concern causing significant financial losses, especially in tropical and subtropical regions of the world. This study aimed at investigating the epidemiologic profile of ticks infesting cattle and molecular identification of R. microplus in the centrally ignored part of Khyber Pakhtunkhwa, Pakistan. A total of 600 cattle from 20 farms were examined for the tick infestation, among them 358 (59.7%) cattle were infested with ticks. A total of 2118 nymph, larvae and adult tick stages were collected and morphologically identified followed by molecular confirmation of Rhipicephalus microplus. Host-based demographic and ecological parameter analysis revealed significantly higher tick infestation in adult, female, exotic, freely grazing, and with irregular/no acaricides treated cattle. The univariate logistic analysis showed that host age, gender, breed, acaricides use, and feeding method were significantly (P < 0.05) associated, whereas multivariate analysis revealed only host breed and feeding method were potential risk factors (P < 0.05) for tick infestation. Microscopy-based examination identified four different species of ticks including R. microplus (44.5%), Hyalomma anatolicum (38.5%), and Hyalomma marginatum (10.5%) and Hyalomma excavatum (6.5%). Tick infestation pattern showed that 55.9% of cattle was found co-infested with R. microplus and H. anatolicum followed by R. microplus and H. anatolicum and H. marginatum (29.3%) then R. microplus, H. anatolicum, H. marginatum, and H. excavatum (11.2%). Sequencing of the second internal transcribed spacer (ITS2-) and 16S rRNA gene fragments also confirmed the molecular identification of Rhipicephalus microplus. Phylogenetic analysis of ITS-2 revealed all sequences clustered in single clade of the R. microplus while the 16S rRNA nucleotide sequences showed that R. microplus in this study was clustered together in clade A along with other isolates from Pakistan, China, and India. The high tick infestation suggests the need for designing strategic and integrated control measures for ticks in order to ensure good health of domestic animals in this region of Pakistan., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Additive manufacturing is an advanced manufacturing manner that seems like the industrial revolution. It has the inborn benefit of producing complex formations, which are distinct from traditional machining technology. Its manufacturing strategy is flexible, including a wide range of materials, and its manufacturing cycle is short. Additive manufacturing techniques are progressively used in bone research and orthopedic operation as more innovative materials are developed. This Review lists the recent research results, analyzes the strengths and weaknesses of diverse three-dimensional printing strategies in orthopedics, and sums up the use of varying 3D printing strategies in surgical guides, surgical implants, surgical predictive models, and bone tissue engineering. Moreover, various postprocessing methods for additive manufacturing for orthopedics are described.

Malignant melanoma remains the life-threatening form of skin cancer with high mortality and poor prognosis. Thus, an ideal melanoma therapeutic strategy is of immediate importance which can remove the primary tumor, as well as inhibit the metastasis and recurrence. Here, we report the fabrication of adjuvant monophosphoryl lipid A (MPLA) lipid bilayer-enveloped and photosensitizer indocyanine green (ICG)-loaded gold nanocages (MLI-AuNCs) for immunogenic phototherapy of aggressive melanoma. Hollow porous AuNCs are used as carriers to deliver MPLA and ICG, and protect ICG from photodegradation. Both AuNCs and ICG absorb near infrared (NIR) light and can be applied in controllable NIR-triggered photothermal and photodynamic combination therapy (PTT/PDT) of melanoma. MLI-AuNCs coated by thermosensitive lipid bilayer exhibit uniform size, good biocompatibility and bioavailability with prominent tumor accumulation, which further improve the PTT/PDT efficacy. MLI-AuNCs under NIR irradiation not only destroy the primary tumor by PTT/PDT, but also elicit robust antitumor immune response with melanoma associated antigens and MPLA released in situ. The released antigens and MPLA subsequently enhance the recruitment and maturation of dendritic cells, which further activate the effector T cells to inhibit metastases and recurrence of melanoma. This immunomodulatory-boosted PTT/PDT nanoplatform provides a new opportunity for highly aggressive melanoma treatment. STATEMENT OF SIGNIFICANCE: An ideal tumor therapeutic strategy not only can remove the primary tumor, but also inhibit metastasis and recurrence. Here, we introduced a versatile nanoplatform MLI-AuNCs for immunogenic phototherapy of aggressive melanoma. Adjuvant MPLA and photosensitizer ICG can be protected and co-delivered to the tumors by thermosensitive lipid-enveloped AuNCs. MLI-AuNCs exhibited prominent tumor accumulation ability and produced the potent PTT/PDT effect to destroy the primary tumors with a single dose of NIR irradiation, as well as elicited the strong antitumor immunity to inhibit the metastasis and relapse. This study may provide a potential therapeutic vaccination strategy against advanced melanoma and other difficult-to-treat cancers., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022. Published by Elsevier Ltd.)

The rapid emergence of resistance to third-generation cephalosporins in Shigella flexneri is crucial in pediatric shigellosis management. Limited studies have been conducted on molecular pattern of antibiotic resistance of S. flexneri in diarrhea endemic areas of Pakistan. The aim of the study was to analyze the antimicrobial resistance of S. flexneri isolated from pediatric diarrheal patients in Peshawar, Pakistan. A total of 199 S. flexneri isolates (clinical, n = 1 55 and non-clinical, n = 44) were investigated for drug resistance and mutational analysis of selected drug resistance genes. All isolates were found to be highly resistant to amoxicillin/clavulanic acid (88%), followed by trimethoprim-sulfamethoxazole (77%), chloramphenicol (43%), and quinolones (41.6%). About 34.5% S. flexneri isolates were found to be resistant to third-generation cephalosporin. None of the isolates was resistant to imipenem, piperacillin-tazobactam, and amikacin. Interestingly high frequency of third-generation cephalosporin resistance was observed in S. flexneri isolated from non-clinical samples (49%) when compared to clinical samples (30.5%). Furthermore, the most prevalent phenotypic-resistant patterns among third-generation cephalosporin-resistant isolates were AMC,CAZ,CPD,CFM,CRO,SXT (13%) followed by OFX,AMC,CAZ,CPD,CFM,CRO,SXT,NA,CIP (10%). The most frequently detected resistance genes were trimethoprim-sulfamethoxazole (sul2 = 84%), beta-lactamase genes (blaOXA = 87%), quinolones (qnrS = 77%), and chloramphenicol (cat = 64%). No mutation was detected in any drug-resistant genes. We are reporting for the first time the sequence of the blaTEM gene in S. flexneri. Furthermore, high third-generation cephalosporin resistance was observed in the patients who practiced self-medication as compared to those who took medication according to physician prescription. This study shows the high emergence of third-generation cephalosporin-resistant S. flexneri isolates, which is a potential threat to the community in the country. This finding will be helpful to develop a suitable antibiotic prescription regime to treat shigellosis., (© 2022. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Highly infectious illnesses caused by pathogens constitute severe threats to public health and lead to global economic loss. The use of robust and programmable clustered regularly interspaced short palindromic repeat and CRISPR-associated protein (CRISPR-Cas) systems, repurposed from genome-engineering applications has markedly improved traditional nucleic acid detection for precise identification, independently enabling rapid diagnostics of multiplex biomarker with genetic and mutation related to tumors, and microbial pathogens. In this review, we delineate the utility of the current CRISPR-Cas enzyme as biosensors by which these effector toolkits achieve recognition, signaling amplification, and finally, accurate detection. Additionally, we discuss the details of the dominance and hurdles related to expanding this revolutionary technology into an effective and convenient contraption crucial for improving the rational redesign to CRISPR/Cas biosensing. Overall, this review provides an insight into the current status of rapid and POC diagnostic systems by CRISPR/Cas tools., (© 2022. The Author(s).)

Molecular hydrogen proved itself as a novel therapeutic candidate and has been thriving from the beginning with its potential clinical significance, higher affinity, and cellular integrity and permeability. Hydrogen Therapy (HT) has gained scientists' attention with the proven clinical ability to attenuate chronic inflammation, diminish oxidative stress, restrict apoptosis, minimize cellular injury, and refine tissue functioning. Therapeutic Implementation of H 2 for disease prevention and treatment is a newly emerging field with limited knowledge available on formulations, tissue-specific effects, efficacy, and safety. This article will discuss HT's therapeutic potential for its efficacy and safety in cardiovascular, respiratory, hematological, metabolic, infectious, and neurodegenerative disorders. In addition to this, the molecular mechanisms and nanotechnological implications of hydrogen therapy will be discussed in detail. Finally, the article will provide insight into advancements and automation, future perspectives, and recommendations. There is a need to study and conduct higher-scale trials targeting personalized treatments under molecular and genetic vitals., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Background: The IL-12/IFN-γ axis plays a vital role in the control of intramacrophagic pathogens including Leishmania infections., Objective: The aim of this study was to investigate genetic defects in the IL-12/IFN-γ axis in cutaneous leishmaniasis patients, using immunological and genetic evaluation., Methods: Enzyme-linked immunosorbent assay was used to quantify IFN-γ , while flow cytometry was performed to analyze surface IL-12Rβ1/IL-12Rβ2 expression and phosphorylation of signal transducers as well as the activator of transcription 4 (pSTAT4). Sequencing was carried out for genetic analysis., Results: The peripheral blood mononuclear cells from the two patients (P1 and P2) demonstrated impaired production of IFN-γ. Furthermore, abolishment of the surface expression of Il-12Rβ1 was observed in lymphocytes, with consequent impairment of STAT4 phosphorylation in the lymphocytes of P1 and P2. IL-12Rβ1 deficiency was identified, which was caused by a novel homozygous missense mutation (c.485>T/p.P162L) and a novel homozygous nonsense mutation (c.805G>T/P.E269*) in the IL-12Rβ2 gene of P1 and P2, respectively. In silico analyses predicted these novel mutations as being pathogenic, causing truncated proteins, with consequent inactivation., Conclusion: Our data have expanded the phenotype and mutation spectra associated with IL-12Rβ1 deficiency, and suggest that patients with CL should be screened for mutations in genes of the IL-12/IFN-γ axis., (Copyright © 2021 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Solar-driven water evaporation is of great importance for freshwater production via solar distillation and has attracted growing attention recently by the development of heat localization strategies. Yet, when polluted water is used as the source water, solar-driven water evaporation might further deteriorate the pollution. In this study, we report the facile preparation of multifunctional Ag 3 PO 4 -reduced graphene oxide (Ag 3 PO 4 -rGO) nanocomposite-coated textiles for clean water production by solar-driven water evaporation, photocatalysis, and disinfection. The multifunctional textiles are obtained through coating Ag 3 PO 4 -rGO nanocomposites onto cotton textile substrates. The resulting textile can float on the water surface, absorb solar light, and convert it into heat, enhancing the water surface temperature and promoting water evaporation. We show that with Ag 3 PO 4 -rGO nanocomposite-coated textiles on the water surface, a high water evaporation rate of 1.31 kg/(m 2 h) can be reached under solar light irradiation. Furthermore, the textiles can simultaneously decompose organic dyes and disinfect pathogenic microbes in water, purifying the raw water during solar-driven water evaporation. Such an all-in-one multifunctional textile provides a facile yet sustainable strategy for freshwater production.

The research work was arranged to check the role of AgNPs and silver ions on callus cells of sugarcane ( Saccharum spp. cv CP-77,400). AgNPs were synthesized chemically and characterized by UV-Vis spectra, XRD and SEM. AgNPs and silver ions were applied in various concentrations (0, 20, 40, 60 ppm) to sugarcane calli and the induced stress was characterized by studying various morphological and biochemical parameters. AgNPs and silver ions treatments produced high levels of malondialdehyde, proline, proteins, TP and TF contents. Similarly, CAT, SOD and POX activity was also significant in both treatments. The lower concentration of AgNPs and silver ions (20 ppm) provided maximum intracellular GSH level. This work mainly showed effects of AgNPs and silver ions on sugarcane calli in terms of morphological aberrations and cell membrane damage due to severe oxidative stress and production of enhanced levels of enzymatic and non-enzymatic antioxidants as self-defence to tolerate oxidative stress by scavenging reactive oxygen species. These preliminary findings will provide the way to study ecotoxicity mechanism of the metal ions and NPs in medicine industry and in vitro toxicity research. Furthermore, silver ions alone and their chemically synthesised AgNPs can be used for various biomedical applications in future.

The interleukin (IL)-12/interferon(IFN)γ axis plays an important role in the control of mycobacterial diseases as demonstrated by the increased susceptibility to mycobacterial species in patients with an inborn error of the IL-12-dependent IFNγ immunity. Here, we report a novel mutation in the IL-12Rβ1 gene in a female Pakistani patient who was born in a consanguineous marriage and developed severe bacille Calmette-Guérin (BCG) infection and recurrent tuberculosis. After reviewing the patient's clinical records, she was investigated for IL-12/IFNγ defects using enzyme-linked immunosorbent assay (ELISA), flow cytometry, and DNA genetic Sanger sequencing. Quantification of secretory cytokines from the patient's peripheral blood mononuclear cells (PBMCs) revealed significantly reduced IFNγ production. Flow cytometric analysis revealed no surface expression of IL-12Rβ1 on PHA-activated T lymphocytes. In addition, IL-12-induced impaired STAT4 phosphorylation in the patient's lymphocytes when compared with those from five healthy controls. The genetic analysis of IL-12Rβ1 gene identified a novel nonsense mutation c.199G>T/p.E67* within exon 3, which encodes part of the cytokine-binding region (CBR). In silico analysis indicates that this novel nonsense mutation generates a truncated protein with an apparent inactivating effect. Our data expand the genetic spectrum of IL-12Rβ1 deficiency. Moreover, our findings highlight the need for developing newborn screening for patients with primary immunodeficiency associated with mycobacterial infections in areas where BCG vaccination is mandatory in order to improve the treatment of patients, and consequently their quality of life.

The experimental work presented in this study was carried out with the hypothesis that plant derived smoke enhanced the morphological, physiological and biochemical attributes of a cereal crop, wheat ( Triticum aestivum L.). Furthermore, this study supported the hypothesis that plant derived smoke acts as vegetative growth promoter, inexpensive, rapid and most appropriate eco-friendly bio-fertilizer for sustainable agriculture. Plant derived smoke was generated by burning of plant material (leaf, straws etc) in a specially designed furnace, and seeds were treated with this smoke for different time duration. Four level of plant derived smoke (1 h, 2 h, 3 h and 4 h) along with control were tested on four wheat cultivars in CRD repeated pot experiment. The smoke-related treatments modified number of morphological, physiological and biochemical features of wheat. Compared with the control, aerosol smoke treatment of the seeds significantly improved root length (2.6%), shoot length (7.7%), RFW (0.04%), SFW (0.7%), SDW (0.1%) and leaf area (63.9%). All the smoke-related treatments significantly promoted RWC (17.3%), water potential (1.5%), osmotic potential (1.4%) and MSI (14.6%) whereas a pronounced increase in chlorophyll a (24.9%), chlorophyll b (21.7%) and total chlorophyll contents (15.5%) were recorded in response to aerosol-smoke treatments. Plant derived smoke exposure applied for short time i.e. 1 h & 2 h induced significant results as compared to prolonged PDS exposure (3 h and 4 h). The best results were observed in Pak-13 and Glaxy-13 wheat cultivars. These findings indicated that the plant-derived smoke treatment has a great potential to improve morphological, physiological and biochemical features of wheat crop.

Cholelithiasis is a major cause of morbidity worldwide. The incidence of gall stone in children in Pakistan has not been sufficiently studied and is increasingly being detected: the reason may be a true rise in the incidence or an improvement in diagnosis due to liberal use of diagnostic facilities and thus the increased opportunity to detect disease. A healthy 2 years and 2 months old male child presented to outpatient with history of recurrent attacks of pain abdomen and anorexia. Ultrasonography showed a 0.6 mm stone, inflammation and pericholecystic fluid. Open cholecystectomy was performed under general anaesthesia. A distended gall bladder with multiple very small calculi was removed. Post-operative stay was uneventful and patient was allowed oral feeds after 24 hours.

In the recent years, nanotechnology has attained much attention in the every field of science. The synthesis, characterisation and applications of metallic nanoparticles (MNPs) have become an important branch of nanotechnology. In the current study, MNPs were synthesised through polyols process and applied in vitro to study their effect on medicinally important plant : Artemisia absinthium. The current study strives to check the effect of MNPs, i.e., Ag, Cu and Au on seed germination, root and shoot length, seedling vigour index (SVI) and biochemical profiling in A. absinthium. The seeds were inoculated on MS medium supplemented with various combinations of MNPs suspension. The seed germination was greatly influenced upon the application of MNPs and was recorded highest for the silver nanoparticles (AgNPs) suspensions. The best result for seed germination (98.6%) was obtained in MS medium supplemented with AgNPs as compared to control (92.9%) and other nanoparticles, i.e., copper (69.6%) and gold (56.5%), respectively, after 35 days of inoculation. Significant results were obtained for root length, shoot length and SVI in response to application of AgNPs as compared to copper nanoparticles (CuNPs) and gold nanoparticles (AuNPs). These nanoparticles (NPs) could induce stress in plants by deploying the endogenous mechanism. In response to these stresses, plants produce various defence compounds. Total phenolic content (TPC) and total flavonoid content (TFC) were significant in the MS medium supplemented with AgNPs as compared to other NPs, while DPPH radical scavenging assay (RSA) was highest in AuNPs treated plantlets. The MNPs showed higher toxicity level and enhanced secondary metabolites production, total phenolic content, total flavonoid content, antioxidant activity, superoxide dismutase (SOD) activity and total protein content.